Rat Gastrin ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Bring all reagents to room temperature (18-25°C) before use for 30min. -Prepare fresh standard for each assay. Use within 4 hours and discard after use. -Centrifuge the sample again after thawing before the assay.	- To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. -Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. -TMB Substrate should remain colorless or light blue until added to the plate. Please protect it from light. -Stop Solution should be added to the plate in the same order as the TMB Substrate.

Upstream tips
-Bring all reagents to room temperature (18-25°C) before use for 30min. -Prepare fresh standard for each assay. Use within 4 hours and discard after use. -Centrifuge the sample again after thawing before the assay.
Protocol tips
- To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. -Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. -TMB Substrate should remain colorless or light blue until added to the plate. Please protect it from light. -Stop Solution should be added to the plate in the same order as the TMB Substrate.

Publication protocol

CCK, MTL, SS and GAS were quantified using ELISA kits (Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's instructions. Briefly, the tissue of transverse colons (100 mg) were homogenized in ice-cold 1x PBS (pH 7.2–7.4) using a glass homogenizer (Sigma-Aldrich Co.). The tissue lysates were then centrifuged at 1,000 rpm for 5 min at 4℃, after which the supernatant was collected for analysis. After addition of the four specific hormone antibodies, separately in each well, the supernatant was incubated for 60 min at 37℃, following which HRP-Streptavidin solution was added and incubated for 60 min at 37℃. The TMP One-Step Substrate Reagent was then added, and the mixture was further incubated for 30 min at 37℃. The reaction was terminated following addition of the stop solution. Finally, the absorbance of the reaction mixture was read at 450 nm using a Molecular Devices VERSA max Plate reader (Sunnyvale, CA, USA).

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Manufacturer protocol

Download the product protocol from Cusabio for Rat Gastrin ELISA Kit below.

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